LOCUS       sequence-295475-        7835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7835)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133834
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             896..916
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1265..1282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1375..1496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1386..1405)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1440..1459
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1611..1657
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1687..1823
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1868..1939
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2165..2753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2245..2264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3253..4462
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4299..4318
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4466..5494
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5521..5569
                     /note="synthetic polyadenylation signal"
     CDS             join(5835..5903,5904..6695)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6053..6072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6708..6844
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6989..7060
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7065..7111)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7129..7177
                     /note="synthetic polyadenylation signal"
     enhancer        7258..7561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7563..7765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7716..7736
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt caagcgtggc aaccgggtgt tgctatgcct aatctttaca aaatgcaaag
       61 aatgctatta gaaaagtgtg accttcaaaa ttatggtgat agtgcaacat tacctaaagg
      121 cataatgatg aatgtcgcaa aatatactca actgtgtcaa tatttaaaca cattaacatt
      181 agctgtaccc tataatatga gagttataca ttttggtgct ggttctgata aaggagttgc
      241 accaggtaca gctgttttaa gacagtggtt gcctacgggt acgctgcttg tcgattcaga
      301 tcttaatgac tttgtctctg atgcagattc aactttgatt ggtgattgtg caactgtaca
      361 tacagctaat aaatgggatc tcattattag tgatatgtac gaccctaaga ctaaaaatgt
      421 tacaaaagaa aatgactcta aagagggttt tttcacttac atttgtgggt ttatacaaca
      481 aaagctagct cttggaggtt ccgtggctat aaagataaca gaacattctt ggaatgctga
      541 tctttataag ctcatgggac acttcgcatg gtggacagcc tttgttacta atgtgaatgc
      601 gtcatcatct gaagcatttt taattggatg taattatctt ggcaaaccac gcgaacaaat
      661 agatggttat gtcatgcatg caaattacat attttggagg aatacaaatc caattcagtt
      721 gtcttcctat tctttatttg acatgagtaa atttcccctt aaattaaggg gtactgctgt
      781 tatgtcttta aaagaaggtc aaatcaatga tatgatttta tctcttctta gtaaaggtag
      841 acttataatt agagaaaaca acagagttgt tatttctagt gatgttcttg ttaacgaaaa
      901 cctttacttt cagtcgatga gtgacaaaat cattcatcta accgatgact cctttgatac
      961 agacgtgttg aaggccgatg gtgcgatact ggtagatttc tgggctgaat ggtgtggccc
     1021 gtgcaagatg atcgcaccca ttctggacga gatcgccgac gagtatcaag gaaaattaac
     1081 tgtcgctaag cttaatatag atcagaaccc agggacggca cctaaatacg gcatcagggg
     1141 aatacctacc ctcctgctct ttaagaatgg ggaagttgcc gcgacaaaag tgggagccct
     1201 gtctaagggc cagctaaaag agttcttgga cgctaacctg gcaggttcag gctcggggca
     1261 catgcaccat caccatcacc atagcagcgg gctcgtgcca cgcggctaag gatccgcagg
     1321 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     1381 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1441 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1501 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1561 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     1621 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     1681 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     1741 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     1801 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     1861 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     1921 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     1981 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     2041 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     2101 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2161 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2221 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2281 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2341 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2401 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2461 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2521 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2581 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2641 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2701 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2761 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2821 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2881 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2941 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     3001 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     3061 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     3121 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     3181 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     3241 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     3301 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     3361 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     3421 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     3481 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     3541 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     3601 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     3661 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     3721 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     3781 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     3841 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3901 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     3961 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     4021 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     4081 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     4141 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     4201 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     4261 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     4321 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     4381 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     4441 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     4501 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4561 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4621 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     4681 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     4741 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4801 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     4861 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4921 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4981 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     5041 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     5101 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     5161 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     5221 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     5281 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     5341 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     5401 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     5461 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     5521 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     5581 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     5641 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     5701 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     5761 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     5821 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5881 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5941 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6001 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6061 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6121 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     6181 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6241 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6301 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6361 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     6421 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     6481 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6541 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6601 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6661 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     6721 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     6781 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6841 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6901 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     6961 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     7021 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     7081 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     7141 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     7201 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     7261 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     7321 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     7381 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     7441 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     7501 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     7561 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     7621 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     7681 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     7741 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     7801 cctaccatcc actcgacaca cccgccagcg gccgc
//
